Quinazolin-4(3h)-ones, methods of making, compositions, and uses

Abstract

The application discloses a quinazoline-4(3H)-ketone compound and a preparation method, a pharmaceutical composition and application thereof, the structure of the compound is as shown in the formula (I), and the compound comprises isomers, pharmaceutically acceptable salts or mixtures thereof. The compound and the pharmaceutical composition thereof have a high inhibitory effect on PARP7, the inhibitory activity reaches a nanomolar concentration level, can significantly promote the release of immune factors, are used for immunotherapy of tumors, and can exert good drug efficacy at a molecular level and a cell level. In addition, the preparation method of the compound is simple and easy to implement.

Description

Technical Field

[0001] This invention relates to a quinazoline-4(3H)-one compound, its preparation method, pharmaceutical composition and application, and more particularly to a quinazoline-4(3H)-one compound that can be prepared as a PARP7 inhibitor drug, has antitumor activity, its preparation method, pharmaceutical composition and application. Background Technology

[0002] Most members of the PARP family in the human body exhibit monoADP ribotransferase activity. The monoPARP protein family is closely related to the occurrence and development of cancer, inflammation, and neurodegenerative diseases. PARP7, a member of the monoPARP protein family, is a novel negative regulator of nucleic acid sensors in cells and is overexpressed in various tumor cells. Because cancer cells can use PARP-7 to inhibit interferon signaling, allowing them to "hide" outside the immune system, many cancer cells depend on PARP-7 for survival. Studies have found that inhibiting PARP7 can restore intracellular interferon signaling, restore the body's innate and adaptive immunity, and thus inhibit cancer cell growth. In cancer models such as lung cancer and colorectal cancer, PARP7 inhibitors have shown durable tumor growth inhibition. Currently, no PARP-7 inhibitors have been approved for marketing. RBN-2397, developed by Ribon, is the first compound with strong inhibitory activity against PARP-7 and is currently in Phase I clinical trials (NCT04053673). Since there are currently no other structural types of inhibitors for this target, designing and synthesizing a series of novel PARP7 inhibitors with novel structural types and evaluating their bioactivity will provide a foundation for further research on this target. Summary of the Invention

[0003] Purpose of the invention: In view of the problems of limited structural types and limited activity of existing compounds, the present invention aims to provide a quinazoline-4(3H)-one compound with excellent antitumor activity, its preparation method, pharmaceutical composition and application.

[0004] Technical solution: As a first aspect of the present invention, the quinazoline-4(3H)-one compounds of the present invention have the structure of formula (I), said compounds comprising isomers thereof, pharmaceutically acceptable salts thereof, or mixtures thereof:

[0005]

[0006] in:

[0007] n is selected from 0, 1, 2, 3, 4 or 5;

[0008] m is selected from 0 or 1;

[0009] R 1 Selected from hydrogen, halogen, cyano, trifluoromethyl, C1-C6 alkyl, C1-C6 alkoxy, methylthio, methanesulfonyl or carbamoyl;

[0010] R 2 Or R 3 Each is independently selected from hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl, heterocycloalkyl, cyano, halogen, difluoromethyl or trifluoromethyl, or R 2 and R 3 Together with the attached carbon atom, it forms a C3-C6 cycloalkyl group; the C3-C6 cycloalkyl group is substituted by one or more of the following groups: hydrogen, methyl, trifluoromethyl, 2,2-difluoroethyl, methoxy, halogen, cyano, amino, methylamino, dimethylamino, diethylamino, acetamino, hydroxy, acetoxy, carboxyl or methoxycarbonyl.

[0011] R 4 It is selected from aryl, heteroaryl or 1,3-benzodioxane, wherein the heteroaryl or 1,3-benzodioxane is substituted by one or more of the following groups: hydrogen, halogen, cyano, trifluoromethyl, 2,2,-difluoroethyl, C1-C6 alkyl, C1-C6 alkoxy, hydroxyl, methoxy, amino, methylamino, dimethylamino, acetamino, carboxyl, methoxycarbonyl or nitro;

[0012] R 5 Selected from hydrogen, cyano, trifluoromethyl, C1-C6 alkyl, C1-C6 alkoxy, or methanesulfonyl;

[0013] A 1 Selected from -NH-, -O-, -S- or -N(CH3)-;

[0014] A 2 The group is selected from -NH-, -O-, -CH2-, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, or 5-8 membered aromatic ring or heteroaromatic ring, wherein any position of the C3-C6 cycloalkyl, heterocycloalkyl, or 5-8 membered aromatic ring or heteroaromatic ring is substituted by one or more of the following groups: hydrogen, halogen, methyl, ethyl, isopropyl, difluoromethyl, difluoromethanesulfonyl, trifluoromethyl, trifluoromethanesulfonyl, methanesulfonyl, cyano, hydroxyl, amino, methylamino, dimethylamino, diethylamino, acetamino, formamide, nitro, methoxy, or ethoxy;

[0015] A 3 Selected from: Among them, R 6 R 7 Or R 8Each group is independently selected from hydrogen, methyl, trifluoromethyl, cyano, hydroxy, methoxy, amino, methylamino, dimethylamino, acetamino, carboxyl, or methoxycarbonyl.

[0016] Preferably, in the above structure:

[0017] n is selected from 0, 1, 2, 3 or 4;

[0018] R 1 Selected from hydrogen or halogen;

[0019] R 2 Or R 3 Each is independently selected from hydrogen, methyl, fluorine, or ethyl; when R 2 R 3 At the same time, with R 2 R 3 The bonded carbon atoms are in racemic, R, or S configurations;

[0020] R 4 Selected from: Among them, Y 1 and Y 2 Each can independently represent N, CH, or CH-R. 9 R 9 Selected from hydrogen, trifluoromethyl, methyl, fluorine, chlorine, bromine, cyano, methoxy, methanesulfonyl, 2,2-difluoroethyl or 4-trifluoromethylphenyl;

[0021] R 5 Selected from hydrogen, methyl, or trifluoromethyl;

[0022] A 1 Selected from -NH- or -O-;

[0023] A 2 Selected from: -NH-, -CH2-, Among them, X 1 X 2 or X 3 Each is independently selected from CH or N, R 10 It is selected from one or more hydrogen, halogen, methyl, ethyl, isopropyl, difluoromethyl, difluoromethanesulfonyl, trifluoromethyl, trifluoromethanesulfonyl, methanesulfonyl, cyano, hydroxy, amino, methylamino, dimethylamino, diethylamino, acetamino, formamide, nitro, methoxy, or ethoxy;

[0024] A 3 Selected from:

[0025] Further optimization, in the above structure:

[0026] R 1Selected from hydrogen or fluorine;

[0027] R 2 Or R 3 Each is independently selected from hydrogen or methyl;

[0028] A 2 Selected from:

[0029] A 3 Selected from:

[0030] Furthermore, in the above structure:

[0031] R 4 Selected from:

[0032] Most preferably, the above-mentioned quinazoline-4(3H)-one compounds are selected from any of the following compounds:

[0033]

[0034]

[0035]

[0036] Pharmaceutically acceptable salts of the above-mentioned quinazoline-4(3H)-one compounds are salts formed by the above-mentioned compounds with an acid selected from hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, carbonic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, malic acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid, or ferulic acid.

[0037] As a second aspect of the present invention, the method for preparing the above-mentioned quinazoline-4(3H)-one compounds is as follows:

[0038] Compound (II) and compound (III) were subjected to substitution, hydrolysis, acylation and deprotection reactions to give compound (I);

[0039]

[0040]

[0041] Where m, n, A 1 A 2 Y 1 Y 2 R 1 R 2 R 3 R 5 R 7 R8 R 10 The definition is as stated above;

[0042] Specifically, compound IV is prepared from compound II by dissolving II and III in a solvent and adding an acid-binding agent to carry out a substitution reaction. The reaction solvent is N,N-dimethylpropenylurea (DMPU), N,N-dimethylformamide (DMF), N,N-dimethylacetamide, tetrahydrofuran (THF), 1,4-dioxane, ethylene glycol dimethyl ether, or acetonitrile, preferably DMPU; the acid-binding agent is sodium carbonate, potassium carbonate, cesium carbonate, triethylamine, or N,N-diisopropylethylamine (DIPEA), preferably cesium carbonate.

[0043] Compound V is prepared from compound IV by dissolving IV in a solvent and then hydrolyzing it in an aqueous solution of an alkali. The reaction solvent is THF, methanol, acetonitrile, or a mixture of any two, preferably a mixture of THF and methanol; the alkali is sodium hydroxide, lithium hydroxide, or potassium hydroxide, preferably sodium hydroxide.

[0044] Compound VII is prepared from compound V by dissolving V in a solvent, adding a condensing agent, and then adding a base and compound VI to carry out a condensation reaction. The solvent is dichloromethane, THF, DMF, 1,4-dioxane, ethylene glycol dimethyl ether, or acetonitrile, preferably DMF; the condensing agent is selected from N,N'-carbonyldiimidazolium (CDI), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI), 1-hydroxybenzotriazole (HOBT), N,N'-dicyclohexylcarbodiimide (DCC), N,N'-diisopropylcarbodiimide (DIC), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU), benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate (HBTU), and benzotriazole-1-yl-oxytripyrrolidine hexafluorophosphate (PyBop), preferably EDCI or HOBT; the base is triethylamine, sodium carbonate, potassium carbonate, or DIPEA, preferably DIPEA.

[0045] Compound I is prepared from compound VII by dissolving VII in a solvent and adding trifluoromethanesulfonic acid to carry out a deprotection reaction. The reaction solvent can be trifluoroacetic acid, acetic acid, THF, methanol, acetonitrile, or any mixture of two solvents, preferably trifluoroacetic acid.

[0046] The corresponding acid is salted with the compound (I) prepared by the above method to obtain a pharmaceutically acceptable salt of the compound.

[0047] As a third aspect of the present invention, the above-mentioned quinazoline-4(3H)-one compounds and pharmaceutically acceptable carriers form a pharmaceutical composition.

[0048] Specifically, the aforementioned quinazoline-4(3H)-one compounds can be added to pharmaceutically acceptable carriers to form common pharmaceutical preparations, such as tablets, capsules, syrups, suspensions, or injections. The preparations can be supplemented with commonly used pharmaceutical excipients such as flavorings, sweeteners, liquid / solid fillers, and diluents.

[0049] As a fourth aspect of the present invention, the above-mentioned quinazoline-4(3H)-one compounds and their pharmaceutical compositions can be prepared as PARP7 inhibitor drugs; more specifically, as antitumor drugs, they are used to treat cancers such as lung squamous cell carcinoma, colon cancer, and breast cancer.

[0050] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages:

[0051] (1) These compounds can effectively inhibit PARP7 enzyme activity, with an enzyme inhibition IC50 value of 100%. 50 The values ​​all reached nanomolar concentration levels, with the optimal value being less than 100 nM; at the same time, it can also significantly promote the release of immune factors, which can be used for the immunotherapy of tumors.

[0052] (2) These compounds and their drug compositions have a wide range of applications and can be prepared as anti-tumor drugs, exerting their effects at the molecular and cellular levels.

[0053] (3) The compound preparation method is simple and feasible. Attached Figure Description

[0054] Figure 1 This is the result of the compound of the present invention promoting the release of interferon. Detailed Implementation

[0055] The technical solution of the present invention will be further described below with reference to the embodiments.

[0056] Example 1: Synthesis of 2-methyl-8-((6-oxo-6-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)hexyl)amino)quinazolin-4(3H)-one (I-1)

[0057] Synthesis of 2-methyl-8-nitroquinazolin-4(3H)-one (II-1-1)

[0058] Add 5.00 g (27.45 mmol) of 2-amino-3-nitrobenzoic acid and 50.0 mL of acetic anhydride to a 100 mL reaction flask and stir to obtain a yellow turbid solution. Heat the solution to 80 °C and maintain the temperature for 2 hours. Monitor the reaction of the starting materials by thin-layer chromatography (V petroleum ether:V ethyl acetate = 1:1) to ensure complete reaction. Stop heating and cool to room temperature. Add 150.0 mL of ice water to quench the reaction solution. Extract with ethyl acetate (50 mL × 3). Combine the organic phases and wash with saturated sodium chloride aqueous solution (100 mL × 3). Dry with anhydrous sodium sulfate and concentrate under reduced pressure to obtain a yellow solid residue. Add 50.0 mL of 25%–28% ammonia solution to the obtained solid residue to obtain a slightly turbid yellow solution. Continue stirring the reaction at room temperature for 1 hour. Monitor the reaction of the starting materials by TLC (V petroleum ether:V ethyl acetate = 1:1) to ensure complete reaction. Stop the reaction, filter, wash the filter cake with water (30 mL × 3), and dry under vacuum to obtain a white solid (II-1-1). Extract the filtrate with ethyl acetate (30 mL × 3), dry the organic phase with anhydrous sodium sulfate, and concentrate under reduced pressure to obtain a yellowish-white solid (II-1-1). Combine the solutions to obtain 4.41 g of (II-1-1), with a yield of 78.29%. mp 268–270℃.

[0059] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 12.36 (s, 1H), 8.30 (dd, J = 8.0, 1.5Hz, 1H), 8.23 ​​(dd, J = 7.8, 1.5Hz, 1H), 7.59 (t, J = 7.9Hz, 1H), 2.38 (s, 3H).

[0060] Synthesis of 3-(4-methoxybenzyl)-2-methyl-8-nitroquinazolin-4(3H)-one (II-1-2)

[0061] Compound II-1-1 (3.74 g, 18.24 mmol), PMB-Cl (3.43 g, 21.89 mmol), and potassium carbonate (3.78 g, 27.36 mmol) were added to a 100 mL reaction flask. Then, 50.0 mL of DMF was added and stirred to obtain a yellow turbid solution. The mixture was heated to 60 °C and reacted for 2 hours. Thin-layer chromatography (V petroleum ether:V ethyl acetate = 1:1) was used to monitor the completeness of the reaction. Heating was stopped, and the mixture was cooled to room temperature. 150 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (50 mL × 3). The organic phase was washed with saturated sodium chloride aqueous solution (100 mL × 3), dried over anhydrous sodium sulfate, and the crude product was purified by silica gel column chromatography (V petroleum ether:V ethyl acetate = 5:1) to give 5.48 g of white solid (II-1-2), yield 92.36%. mp 118–120 °C.

[0062] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.38 (dd, J=8.0, 1.5Hz, 1H), 8.29 (dd, J=7.8, 1.5Hz, 1H), 7.65 (t, J =7.9Hz,1H),7.20(d,J=8.7Hz,2H),6.91(d,J=8.7Hz,2H),5.31(s,2H),3.73(s,3H),2.52(s,3H).

[0063] Synthesis of 8-amino-3-(4-methoxybenzyl)-2-methylquinazolin-4(3H)-one (II-1)

[0064] Add 6.05 g (18.60 mmol) of II-1-2 and stannous chloride dihydrate (25.18 g, 111.60 mmol) to a 100 mL reaction flask, then add 50.0 mL of ethyl acetate and stir to obtain a clear yellow solution. Heat to 70 °C and maintain the temperature for 1 hour. Monitor the reaction of the starting materials by thin-layer chromatography (V petroleum ether:V ethyl acetate = 1:1) to ensure complete reaction. Stop heating and cool to room temperature. Add sodium hydroxide solution to the reaction solution to adjust the pH to 13-14, then extract with ethyl acetate (50 mL × 3). Wash the organic phase with saturated sodium chloride aqueous solution (100 mL × 3), dry with anhydrous sodium sulfate, and purify the crude product by silica gel column chromatography (V petroleum ether:V ethyl acetate = 3:1) to obtain 4.66 g of white solid (II-1), yield 84.83%. mp 132-134 °C.

[0065] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 7.27 (dd, J=7.9, 1.5Hz, 1H), 7.19 (t, J=7.7Hz, 1H), 7.14 (d, J=8.6Hz, 2H) ,6.97(dd,J=7.7,1.5Hz,1H),6.90(d,J=8.6Hz,2H),5.65(s,2H),5.28(s,2H),3.72(s,3H),2.51(s,3H).

[0066] Synthesis of methyl 6-((3-(4-methoxybenzyl)-2-methyl-4-oxo-3,4-dihydroquinazolin-8-yl)amino)hexanoate (IV-1)

[0067] Add 8-amino-3-(4-methoxybenzyl)-2-methylquinazoline-4(3H)-one (II-1) (0.60 g, 2.03 mmol), methyl 6-bromohexanoate (III-1) (1.06 g, 5.08 mmol), cesium carbonate (1.3 g, 4.06 mmol), and sodium iodide (0.31 g, 2.03 mmol) to a 25 mL reaction flask, then add DMF. 10.0 mL of the solution was stirred to obtain a yellow turbid solution. The temperature was raised to 105℃ and the reaction was carried out for 5 hours. Thin-layer chromatography (V petroleum ether:V ethyl acetate = 1:1) was used to monitor the completeness of the reaction. Heating was stopped, and the solution was cooled to room temperature. 30 mL of water was added to the reaction solution, and then extracted with ethyl acetate (20 mL × 3). The organic phase was washed with saturated sodium chloride aqueous solution (50 mL × 3), dried over anhydrous sodium sulfate, and the crude product was purified by silica gel column chromatography (V petroleum ether:V ethyl acetate = 8:1) to obtain 0.65 g of yellow oil (IV-1), with a yield of 75.90%.

[0068] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 7.29–7.22 (m, 2H), 7.13 (d, J = 8.7Hz, 2H), 6.90(d,J=8.7Hz,2H),6.81(dd,J=6.2,3.1Hz,1H),5.92(t,J=5.9Hz,1H),5.29(s,2H),3 .72(s,3H),3.57(s,3H),3.18(q,J=6.6Hz,2H),2.51(s,3H),2.30(q,J=7.8Hz,2H),1.63 –1.55(m,4H),1.42–1.36(m,2H).

[0069] Synthesis of 6-((3-(4-methoxybenzyl)-2-methyl-4-oxo-3,4-dihydroquinazolin-8-yl)amino)hexanoic acid (V-1)

[0070] Compound IV-1 (0.62 g, 1.46 mmol) and sodium hydroxide (0.16 g, 3.92 mmol) were added to a 25 mL reaction flask, followed by 1.0 mL of methanol, 1.0 mL of water, and 3.0 mL of THF. The mixture was stirred to obtain a white, turbid solution. The solution was heated to 50 °C and maintained at this temperature for 0.5 hours, yielding a colorless, clear solution. Thin-layer chromatography (V petroleum ether:V ethyl acetate = 1:1) was used to monitor the completeness of the reaction. Heating was stopped, and the mixture was cooled to room temperature. 10 mL of water was added to the reaction solution, followed by extraction with ethyl acetate (10 mL × 3). The organic phase was discarded. 2.0 mL of 2M dilute hydrochloric acid solution was added to the aqueous phase to adjust the pH to acidic. The phase was then extracted with ethyl acetate (10 mL × 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 0.55 g of a white solid (V-1), with a yield of 91.75%. (mp 124–126 °C)

[0071] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 12.01 (s, 1H), 7.29–7.22 (m, 2H), 7.14 (d, J = 8.7Hz, 2H), 6.90 (d, J = 8.7Hz, 2H), 6.81 (dd, J = 6.3, 2.9Hz, 1H), 5.9 2(t,J=5.9Hz,1H),5.29(s,2H),3.72(s,3H),3.18(q,J=6.7Hz,2H),2. 51(s,3H),2.22(t,J=7.3Hz,2H),1.66–1.50(m,4H),1.43–1.31(m,2H).

[0072] Synthesis of 4-tert-butyl-1-(5-iodopyrimidin-2-yl)piperazine carboxylate (VI-1-1)

[0073] 2-Chloro-5-iodopyrimidine (12.50 g, 51.99 mmol), tert-butyl piperazine-1-carboxylate (11.62 g, 62.39 mmol), and potassium carbonate (8.3 g, 60.14 mmol) were added to a 250 mL reaction flask, followed by 50 mL of N-methylpyrrolidone (NMP). The mixture was heated to 100 °C and reacted for 6 hours. Thin-layer chromatography (V petroleum ether:V ethyl acetate = 8:1) was used to monitor the reaction until complete. 200 mL of water was added, and a white solid precipitated. After thorough stirring, the mixture was filtered, and the filter cake was dried under vacuum to give 19.93 g of a white solid (VI-1-1), with a yield of 98.24%. The mp values ​​were 113–115 °C.

[0074] 1H NMR (300MHz, Chloroform-d) δ (ppm): 8.40 (s, 2H), 3.81–3.71 (m, 4H), 3.53–3.43 (m, 4H), 1.49 (s, 9H).

[0075] Synthesis of 4-(5-trifluoromethylpyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (VI-1-2)

[0076] Add 4-tert-butyl-1-(5-iodopyrimidin-2-yl)piperazine carboxylate (VI-1-1) (23.64 g, 60.58 mmol) and cuprous iodide (23.08 g, 121.16 mmol) to a 500 mL three-necked flask, then add 100 mL of [the solution / formula]. Under nitrogen protection (NMP), methyl 2,2-difluoro-2-(fluorosulfonyl)acetate (23.28 g, 121.16 mmol) was slowly added dropwise at room temperature. After the addition was complete, the mixture was heated to 100 °C and reacted for 8 hours. The reaction was monitored for completeness by thin-layer chromatography (V petroleum ether:V dichloromethane:V methanol = 15:10:2). 200 mL of water was added, and the mixture was extracted with ethyl acetate (100 mL × 3). The combined organic phases were washed with saturated sodium chloride aqueous solution (40 mL × 3), dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by silica gel column chromatography (V dichloromethane:V methanol = 15:1) to give 19.14 g of white solid (VI-1-2), yield 95.06%. mp 125–127 °C.

[0077] 1 H NMR (300MHz, Chloroform-d) δ (ppm): 8.50 (s, 2H), 3.93–3.86 (m, 4H), 3.54–3.48 (m, 4H), 1.49 (s, 9H).

[0078] Synthesis of 2-(piperazin-1-yl)-5-trifluoromethylpyrimidine hydrochloride (VI-1)

[0079] 4-(5-trifluoromethylpyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (VI-1-2) (1.88 g, 5.66 mmol) was added to a 25 mL reaction flask, followed by the addition of 10 mL of saturated HCl in EA solution, yielding a white turbid solution. After stirring at room temperature for 1 hour, the reaction was monitored for completion by thin-layer chromatography (V dichloromethane:V methanol = 15:1). The mixture was filtered, and the filter cake was washed with ethyl acetate (5 mL × 3), dried under vacuum, to give 1.45 g of white solid (VI-1), yield 95.61%. mp > 270 °C.

[0080] 1H NMR (400MHz, DMSO-d6) δ (ppm): 9.71 (s, 2H), 8.79 (s, 2H), 4.12–4.06 (m, 4H), 3.21–3.13 (m, 4H).

[0081] Synthesis of 3-(4-methoxybenzyl)-2-methyl-8-((6-oxo-6-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)hexyl)amino)quinazolin-4(3H)-one (VII-1)

[0082] Compound V-1 (0.15 g, 0.37 mmol), compound VI-1 (0.10 g, 0.37 mmol), EDCI (0.14 g, 0.73 mmol), HOBt (0.1 g, 0.73 mmol), and DIEA (0.19 g, 1.47 mmol) were added to a 10 mL reaction flask. Then, 3.0 mL of DMF was added and the mixture was stirred to obtain a pale yellow, clear solution. The reaction was carried out at room temperature for 12 hours. Thin-layer chromatography (V dichloromethane:V methanol = 15:1) was used to monitor the completeness of the reaction. 5.0 mL of water was added to the reaction solution, followed by extraction with ethyl acetate (5 mL × 3). The organic phase was washed with saturated sodium chloride solution (15 mL × 3), dried over anhydrous sodium sulfate, and the crude product was purified by silica gel column chromatography (V petroleum ether:V ethyl acetate = 1:1) to give 0.14 g of a white solid (VII-1), with a yield of 59.97%. The reaction temperature was mp 135–137 °C.

[0083] 1 H NMR (300MHz, DMSO-d6) δ (ppm): 8.73 (d, J = 0.9Hz, 2H), 7.30–7.19 (m, 2H), 7.13(d,J=8.7Hz,2H),6.90(d,J=8.7Hz,2H),6.82(dd,J=6.9,2.3Hz,1H),5.93(t,J=5 .9Hz,1H),5.28(s,2H),3.87–3.81(m,2H),3.81–3.75(m,2H),3.72(s,3H),3.59–3.51 (m,4H),3.20(q,J=6.6Hz,2H),2.51(s,3H),2.38(t,J=7.3Hz,2H),1.69–1.51(m,4H,),1.46–1.34(m,2H).

[0084] Synthesis of 2-methyl-8-((6-oxo-6-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)hexyl)amino)quinazolin-4(3H)-one (I-1)

[0085] Compound VII-1 (96.00 mg, 0.15 mmol) was added to a 10 mL reaction flask, followed by 3.0 mL of TFA and 0.3 mL of TfOH, and stirred to obtain a clear, purple-red solution. The reaction was carried out at room temperature for 3 hours, and the reaction was monitored for completeness by thin-layer chromatography (V dichloromethane: V methanol = 15:1). The reaction was stopped, and the pH was adjusted to alkaline by adding saturated sodium bicarbonate solution to the reaction solution. The solution was then extracted with ethyl acetate (10 mL × 3), and the organic phase was washed with saturated sodium chloride aqueous solution (10 mL × 3), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (V petroleum ether: V ethyl acetate = 1:2) to give 53.00 mg of white solid (I-1), with a yield of 68.38%. mp 199–201 °C.

[0086] 1 H NMR(300MHz,DMSO-d6)δ(ppm):12.11(s,1H-),8.73(s,2H),7.23–7.13(m, 2H),6.78(dd,J=7.1,2.2Hz,1H),5.84(t,J=5.8Hz,1H),3.87–3.82(m,2H),3.81–3.77 (m,2H),3.58–3.53(m,4H),3.18(q,J=6.6Hz,2H),2.39(t,J=7.3Hz,2H),2.35(s,3H), 1.68–1.53(m,4H),1.46–1.35(m,2H).

[0087] 1 H NMR(300MHz,DMSO-d6,D2O)δ(ppm):8.71(s,2H),7.27–7.14(m,2H),6.81 (dd,J=7.4,1.9Hz,1H),3.86–3.81(m,2H),3.81–3.76(m,2H),3.59–3.52(m,4H),3.18 (t,J=7.0Hz,2H),2.40(t,J=7.2Hz,2H),2.36(s,3H),1.70–1.52(m,4H),1.47–1.35(m, 2H).

[0088] 13C NMR (101MHz, DMSO) δ (ppm): 171.34, 162.47, 162.24, 156.26 (q, J = 3.5Hz), 152.08, 144.09, 136.70, 126.99, 124.86 (q, J = 271. 0Hz), 120.76, 112.24 (q, J = 33.2Hz), 110.99, 110.93, 44.87, 44.10, 43.77, 42.91, 41.04, 32.71, 28.87, 26.85, 25.02, 21.93.

[0089] HRMS(ESI):m / z[M+H] + Calcd for C 24 H 28 F3N7O2:504.2335,Found:504.2325.

[0090] Following the preparation method of Example 1, the following compounds were prepared:

[0091]

[0092]

[0093] Example 2: Inhibitory activity of the compound against PARP7

[0094] Experimental materials: PARP7 Chemiluminescent Assay Kit, BPS Bioscience; DMSO, Sinopharm, Nivo, PerkinElmer.

[0095] Experimental methods:

[0096] (1) Preparation of solutions and buffer solutions:

[0097] Preparation of 10X PBS: Weigh 720 mg KH2PO4, 45 g NaCl and 5.311 g Na2HPO4·12H2O and dissolve them in 500 mL of deionized water. Adjust the pH of the system to 7.4, sterilize at 121 °C for 30 min, cool and store at 4 °C for later use.

[0098] Preparation of 1X PBS: Dilute 10X PBS with deionized water 10 times, that is, add 1 part of 10X PBS to 9 parts of deionized water for dilution.

[0099] Wash buffer preparation: 1X PBS contains 0.05% Tween-20.

[0100] Preparation of 1X PARP buffer: (Prepare fresh for immediate use) Dilute 10X PARP buffer 10 times with deionized water and place on ice for later use.

[0101] (2) Preparation of the working solution concentration of the compound:

[0102] According to the testing requirements, the compound to be tested is diluted with 100% DMSO to the required concentration, and then diluted 10 times with 1X PARP buffer to prepare a 10X compound working solution.

[0103] (3) Experimental steps:

[0104] a. Thaw the 5X histone mixture on ice the day before the experiment;

[0105] b. Preparation of 1X histone mixture: Prepare 1X histone mixture by mixing 5X histone mixture with 1X PBS; add 25 μL of 1X histone mixture to each well of the test plate and incubate overnight at 4°C.

[0106] c. Add 100 μL of Blocking buffer to each well into the test plate and incubate at 25 °C for 90 min;

[0107] d. After incubation, shake off the liquid in the test plate and wash the plate three times.

[0108] e. Take 2.5 μL of the compound working solution for each well and add it to the test plate according to the experimental layout diagram; add the corresponding volume of 1X PARP buffer containing 10% DMSO to the positive control well, and add the corresponding volume of 1X PARP buffer to the blank control well.

[0109] f. After the enzyme is completely dissolved, dilute the enzyme stock solution to 6 ng / μL with 1X PARP buffer;

[0110] g. Add 10 μL of enzyme solution per well to the test plate, and add the corresponding volume of 1X PARP buffer to the blank control wells. The enzyme amount is now 60 ng per well. Note: This step must be performed on ice.

[0111] h. Add 12.5 μL of master mixture (12.5 μL master mixture includes 1.25 μL of 10X PARP buffer, 1.25 μL of Opti-PARP 10X Assay mixture and 10 μL of water) to each well of the test plate; seal the test plate and incubate at 25°C for 60 min;

[0112] i. After incubation, shake off the liquid in the test plate and wash the plate three times.

[0113] j. Dilute the Streptavidin-HRP in the kit 50 times with Blocking buffer solution, add 25 μL to each well of the test plate, and incubate at 25 °C for 30 min;

[0114] k. After incubation, shake off the liquid in the test plate and wash the plate three times.

[0115] 1. Mix ELISA ECL Substrate A and ELISA ECL Substrate B in the 1:1 kit, add 50 μL of the mixture to each well of the test plate, and immediately perform Luminescence detection using Nivo to read the luminescence value (RLU).

[0116] m. Enzyme rate calculation:

[0117] %Enzyme Activity = (RLU(Sample) - RLU(Blank)) / (RLU(Pos.Ctrl) - RLU(Blank)) × 100%; Enzyme Inhibition Rate = 1 - %Enzyme Activity. IC50 was performed using Prism GraphPad software. 50 The fitting results are shown in Table 1 below.

[0118] Table 1. Inhibitory activity of the test compounds against PARP7

[0119]

[0120]

[0121] Note: "++" indicates IC. 50 <100nM; "++" means 100nM <IC 50 <500nM.

[0122] As shown in Table 1, the compounds of the present invention all exhibit good inhibitory activity against PARP7 enzyme, IC50. 50The values ​​reached the nanomolar level. Among them, compounds I-4 to I-6 and I-8 showed IC50 values ​​for the effects on PARP7 enzyme activity. 50 The values ​​are all less than 100 nM.

[0123] Example 3: The promoting effect of the compound on interferon release

[0124] The level of interferon-β induced by PARP7 inhibitor in CT-26 cells was determined in the presence of the STING agonist DMXAA. CT-26 cells in logarithmic growth phase were plated in 96-well plates and incubated overnight at 37°C in a 5% CO2 incubator until adherence. Cells were then co-treated with a specific concentration of PARP7 inhibitor and 50 μg / mL DMXAA for 24 h, and the supernatant was collected. Cells were then processed using ELISA (R&D, Mouse IFN--beta DuoSet ELISA) according to the kit instructions. Results are shown below. Figure 1 .Depend on Figure 1 It is evident that the compounds of this invention can significantly promote the release of interferon β, and therefore can be used for the immunotherapy of tumors.

Claims

1. A quinazoline-4(3) H )-ketone compounds, characterized in that, Having the structure of formula (I), the compound comprises a pharmaceutically acceptable salt: （I）， in: n is selected from 0, 1, 2, 3, 4 or 5; m is selected from 0 or 1; R 1 Selected from hydrogen; R 2 Or R 3 Each is independently selected from hydrogen or methyl; R 4 Selected from ; R 5 Selected from methyl; A 1 Selected from -NH-; A 2 Selected from -CH2-; A 3 Selected from .

2. A quinazoline-4(3) H )-ketone compounds, characterized in that, Selected from any of the following compounds: 2-Methyl-8-((6-oxo-6-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)hexyl)amino)quinazolin-4(3 H )-keto(I-1), 2-Methyl-8-((2-oxo-2-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)ethyl)amino)quinazolin-4(3 H )-keto(I-2), 2-Methyl-8-((1-oxo-1-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)propyl-2-yl)amino)quinazolin-4(3 H )-keto(I-3), 2-Methyl-8-((3-oxo-3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)propyl)amino)quinazolin-4(3 H )-keto(I-4), 2-Methyl-8-((4-oxo-4-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)butyl)amino)quinazolin-4(3 H )-keto(I-5), 2-Methyl-8-((5-oxo-5-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazin-1-yl)pentyl)amino)quinazolin-4(3 H )-keto(I-6), 2-Methyl-8-((3-(4-(5-(trifluoromethyl)pyrimidin-2-yl)piperazine-1-carbonyl)benzyl)amino)quinazolin-4(3 H )-keto(I-7), 8-((4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)-4-oxobutyl)amino)-2-methylquinazoline-4(3 H )-keto(I-8), 2-Methyl-8-((4-oxo-4-(4-(5-(trifluoromethyl)pyridin-2-yl)piperazin-1-yl)butyl)amino)quinazolin-4(3 H )-keto(I-9), 2-Methyl-8-((5-(4-(5-methylpyrimidin-2-yl)piperazin-1-yl)-5-oxopentyl)amino)quinazolin-4(3 H )-keto(I-10), 8-((5-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)-5-oxopentyl)amino)-2-methylquinazoline-4(3 H )-keto(I-11), 8-((5-(4-(5-methoxypyrimidin-2-yl)piperazin-1-yl)-5-oxopentyl)amino)-2-methylquinazoline-4(3 H )-keto(I-12), 2-Methyl-8-((5-oxo-5-(4-(pyrimidin-2-yl)piperazin-1-yl)pentyl)amino)quinazolin-4(3) H )-keto(I-13), 2-Methyl-8-((5-oxo-5-(4-(5-(trifluoromethyl)pyridin-2-yl)piperazin-1-yl)pentyl)amino)quinazolin-4(3 H )-keto(I-14), 8-((5-(4-(2,2-difluorobenzo[ d [1,3]dioxacyclopenten-5-yl)piperazin-1-yl)-5-oxopentyl)amino)-2-methylquinazoline-4(3) H )-keto(I-15).

3. The quinazoline-4(3) according to claim 1 or 2 H )-ketone compounds, characterized in that, The pharmaceutically acceptable salt is a salt formed by the compound and an acid selected from hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, carbonic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, malic acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid, or ferulic acid.

4. A quinazoline-4 (3) according to any one of claims 1 to 3 H A method for preparing ketone compounds, characterized in that, The preparation method is as follows: Compound (II) and compound (III) were subjected to substitution, hydrolysis, acylation and deprotection reactions to obtain compound (I); ； Where m, n, A 1 A 2 R 1 R 2 R 3 R 5 The definition is as described in claim 1. The definition of A as in claim 1 3 The above, The definition of R as in claim 1 4 The above; The corresponding acid is salted with the compound (I) prepared by the above method to obtain a pharmaceutically acceptable salt of the compound.

5. A pharmaceutical composition, characterized in that, Contains any one of the quinazoline-4 (3) according to claims 1 to 3 H )-ketone compounds and pharmaceutically acceptable carriers.

6. A quinazoline-4 (3) according to any one of claims 1 to 3 H The use of ketone compounds or the pharmaceutical composition of claim 5 in the preparation of PARP7 inhibitor drugs.

7. The application according to claim 6, characterized in that, The drug in question is an anti-tumor drug.

8. The application according to claim 7, characterized in that, The drug in question is an anti-squamous cell carcinoma of the lung, colon cancer, or breast cancer.

Images